Improved peptide mapping using phytic acid as ion-pairing buffer additive in capillary electrophoresis

Kornfelt, Troels; Vinther, Anders; Okafo, George; Camilleri, Patrick

doi:10.1016/0021-9673(95)01044-0

Cited by 27 publications

(12 citation statements)

References 13 publications

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“…It is worthwhile noting that other multiply charged anions are also known to interact strongly with peptides. An example is PA, which is often applied as a BGE additive for the separation of peptides by CE [38]. Similarly to S-b-CD, the effect of PA on the mobility of peptides is quite strong and even at low PA concentrations the migration of peptides is totally reversed.…”

Section: Effect Of Sulphate and Organic Sulphated And Sulphonated Commentioning

confidence: 99%

CE study of neuroprotective humanin peptide and its derivatives: Interactions with phosphate, sulphate, alkylsulphonates and sulphated‐β‐CD

Havel

Li²,

Macka

2008

Electrophoresis

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Humanin (HN), Met-Ala-Pro-Arg-Gly-Phe-Ser-Cys-Leu-Leu-Leu-Leu-Thr-Ser-Glu-IIe-Asp-Leu-Pro-Val-Lys-Arg-Arg-Ala, recently discovered in the human brain, is an important neuroprotective peptide. Some derivatives of HN show even higher biological activity, for example [G-14]-HN, where Ser at position 14 is replaced with Gly. As structurally related HN peptide derivatives have similar chemical properties, their separation by CE is difficult. In this work, the electrophoretic behaviour of HN derivatives including [G-14]-HN, a tryptophan HN derivative [W-14]-HN, several other HN derivatives and HN fragments was studied. While phosphate buffer was used as the general BGE, the effects of the buffer concentration and various additives were examined, including sulphate, heptane sulphonate, 2-morpholinoethanesulphonic acid N-[tris(hydroxymethyl)methyl]-2-aminoethane sulphonic acid (TES), sulphated-beta-CD and beta-CD. Separation efficiency of 200,000 theoretical plates was achieved in a BGE of 80 mM phosphate at pH 2.5 where seven out of nine major peaks were partially separated. By investigating the influence of concentration of the interrogated ions on peptides migration, the association between positively charged protonated sites of peptides and various anions was proved. Especially a strong interaction with phosphate, sulphate and sulphonate groups was established. Conditional stability constant of the [Pep(z+), (H(2)PO(4)(-))(n)](z - n) ion associate (n = 1) for [G-14]-HN equals to log K approximately 1.78.

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Section: Effect Of Sulphate and Organic Sulphated And Sulphonated Commentioning

confidence: 99%

CE study of neuroprotective humanin peptide and its derivatives: Interactions with phosphate, sulphate, alkylsulphonates and sulphated‐β‐CD

Havel

Li²,

Macka

2008

Electrophoresis

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“…On the other hand, capillary zone electrophoresis (CZE) exploits analyte charge or, more precisely, the mass-to-charge ratio. The value of these two peptide characteristics for peptide analysis in general, and peptide mapping in particular, has been demonstrated many times [17][18][19], with RP-HPLC and CZE also proving to complement each other for an aforementioned multidimensional approach to peptide separations. Indeed, the hydrophobicity-based method (RP-HPLC) and the charge-based method (CZE) can be coupled by coupling the instruments on-line [20,21] or out of line [23,24].…”

Section: Introductionmentioning

confidence: 96%

Capillary electrophoresis of cationic random coil peptide standards: Effect of anionic ion‐pairing reagents and comparison with reversed‐phase chromatography

2004

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The present study compares a charge/hydrophobicity capillary electrophoresis (CE) approach to reversed-phase high-performance liquid chromatography (RP-HPLC) for the separation of three series of four synthetic, random coil peptide standards. Each series has peptides of the same positive charge (+1, +2 and +3 series) and length but differing in hydrophobicity. Complete resolution of the 12 peptides was achieved via a novel CE approach: a capillary zone electrophoresis (CZE) mode effected a separation of identically charged peptides; within each charged group of peptides, the addition of perfluorinated acid anionic ion-pairing reagents allowed resolution of the peptides through a mechanism based on peptide hydrophobicity which we have termed ioninteraction (II)-CZE. The peak capacity and peptide resolution of this CE approach was superior to that of RP-HPLC and stresses an important role for CE for peptide/proteomic applications.

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“…These analyte-wall interactions may in turn lead to band broadening effects (decreased separation efficiency and resolution), variable migration times and poor recovery of the analytes. Numerous strategies have been reported to limit analyte adsorption; such as low/high pH ranges [9,10], high ionic strength [9][10][11], buffer additives [12][13][14] as well as dynamic and static capillary surface derivatizations [15]. Despite improvements, a lot of these strategies are not compatible with MS and especially not with electrospray ionization.…”

Section: Introductionmentioning

confidence: 98%

Monomer surface modifications for rapid peptide analysis by capillary electrophoresis and capillary electrochromatography coupled to electrospray ionization‐mass spectrometry

et al. 2004

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In this study, positively charged alkylaminosilyl monomers were used to modify the inner surface of fused silica capillaries, which subsequently were employed in capillary electrophoresis (CE) and capillary electrochromatography (CEC). The obtained surfaces yield a reversed electroosmotic flow (EOF) and have varying carbon chain lengths, that interact with the analytes and give chromatographic retention. The coating procedure is very simple and fast. The performance of the modified capillaries was evaluated regarding pH influence on EOF and chromatographic interactions. The experiments were conducted with UV and mass spectrometry (MS) and applied to the separation of various neuropeptides. The derivatized surfaces showed a linear (R(2) approximately 0.99) pH dependence with isoelectric points (pI) at 8.6-8.8. Rapid separations of peptide standards and a protein digest with efficiencies as high as 5 x 10(5) plates/m were performed.

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Improved peptide mapping using phytic acid as ion-pairing buffer additive in capillary electrophoresis

Cited by 27 publications

References 13 publications

CE study of neuroprotective humanin peptide and its derivatives: Interactions with phosphate, sulphate, alkylsulphonates and sulphated‐β‐CD

CE study of neuroprotective humanin peptide and its derivatives: Interactions with phosphate, sulphate, alkylsulphonates and sulphated‐β‐CD

Capillary electrophoresis of cationic random coil peptide standards: Effect of anionic ion‐pairing reagents and comparison with reversed‐phase chromatography

Monomer surface modifications for rapid peptide analysis by capillary electrophoresis and capillary electrochromatography coupled to electrospray ionization‐mass spectrometry

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